Breeding Wheat and Triticale for lignocellulosic bioethanol production using basic and molecular

La impronta del ser humano sobre el medio ambiente es hoy una realidad. El problema medioambiental que se cierne sobre la globalidad del planeta se ha visto acelerado por la masiva emisión de gases de efecto invernadero que produce la quema de combustibles fósiles para la obtención de energía. Por otro lado esta la demanda de energía, que se ve incrementada año a año por el constante aumento de la población mundial. Además, a estas tensiones sobre la demanda de energía hay que sumar la escasez de petróleo que se preveé en el corto-medio plazo, pues como sabemos, se trata de una fuente de energía no renovable. Es difícil encontrar alternativas energéticas viables que se adapten a las infraestructuras y necesidades energéticas actuales. Aun así, una de las alternativas más prometedoras para contribuir a paliar estos problemas, es el uso de la biomasa vegetal para la obtención de bioetanol. Pero a pesar de ser una muy buena opción, su uso supone destinar alimentos para la obtención de etanol. Esto provoca que los precios de los alimentos se encarezcan, creando un grave problema de abastecimiento y seguridad alimentaria en las poblaciones más desfavorecidas. El uso de los biocombustibles genera un intenso debate por su efecto en los precios de los alimentos, como también en su posible papel en la mitigación de cambio climático, así como en el desarrollo agrícola. Estos temas de debate fueron tratados en la Conferencia de Alto Nivel sobre Seguridad Alimentaria Mundial: “Los Desafíos del Cambio Climático y la Bioenergía”, donde se evaluó detalladamente la perspectiva futura, riesgos y oportunidades que podrían generar los biocombustibles, y que quedó como tema central del informe de la FAO de 2008 acerca de “El estado mundial de la agricultura y a alimentación”. La introducción del biocombustible de segunda generación, como aquellos que usan biomasa lignocelulósica para la producción de bioetanol, abre una posibilidad a la producción de un carburante menos contaminante que el petróleo, sin entrar en competencia directa por el alimento. La biomasa lignocelulósica para la producción de bioetanol proviene principalmente de residuos agrícolas como restos de poda, rastrojos, restos de madera, paja de maíz, trigo o arroz, bagazo (residuos de la caña de azúcar y el sorgo), etc. Esta biomasa es el principal componente de la pared celular de las plantas, que por sus características químicas y estructurales, es extremadamente resistente a la digestión enzimática necesaria que se pretende hacer para liberar los azúcares atrapados en ella. Tan recalcitrante es esta estructura, que la bioconversión en etanol de esta materia prima hace que el proceso actualmente sea inviable económicamente. El principal objetivo pues, es que el proceso de producción de bioetanol llegue a ser económicamente viable, y para conseguirlo podemos hacerlo a través una digestión más eficiente mediante, o por la obtención de biomasa menos recalcitrantes y más fácilmente accesibles a las enzimas, o ambas. Este trabajo ha abordado este reto mediante una búsqueda de materiales lignocelulósicos más fácilmente accesibles a las enzimas hidrolíticas. Para llevarlo a cabo se han usado herramientas de mejora clásicas y moleculares. Una colección de sesenta y seis genotipos de trigo, triticale y cebada se han caracterizado fenotípicamente a lo largo de su desarrollo. Los genotipos mostraron gran variabilidad para el factor sacarificación, que estuvo correlacionado negativamente con el contenido de lignina en la pared celular. Además, los resultados mostraron que éste y otros factores podrían ser evaluados con cierta precisión usando el tratamiento de imágenes con plataformas aéreas no tripuladas (UAV – unmanned aerial vehicle), antes de ser recolectadas, de forma rápida y no destructiva. Además, varios parentales provenientes de poblaciones de mapeo mostraron diferencias contrastantes para el grado de sacarificación. Por último, se hizo una selección de genes involucrados en la síntesis de componentes de pared y se silenciaron mediante micros-ARN (miARN) en Brachypodium distachyon. Las plantas transgénicas mostraron alteraciones en las células de pared en del tallo y en algunos componentes de pared que la forman, dando lugar a un mayor potencial de sacarificación y alteraciones en los principales componentes de la pared celular.The imprint of the human being on the environment is today a reality. The environmental problem that hangs over the global nature of the planet has been accelerated by the massive emission of greenhouse gases produced by the burning of fossil fuels. On the other hand, this demand for energy increases every year due to the increase in world population. In addition, we must add to these tensions on demand, the shortage of oil that is expected in the short-medium term, as we know, it is a non-renewable source of energy. The infrastructures and energy needs that we have today do not make it easy for us to find economically viable energy alternatives. Even so, one of the most promising alternatives to help alleviate these problems is the use of vegetable biomass for bioethanol production. But this, despite being a very good option, means entering into competition with food production. As a result, large increases in food prices could occur, creating a serious problem of supply and food security in the most disadvantaged populations. The use of biofuels generates an intense debate because of its effect on food prices, as well as its possible role in mitigating climate change, as well as agricultural development. These topics were discussed at the High Level Conference onWorld Food Security: “The Challenges of Climate Change and Bioenergy”, where the future perspective, risks and opportunities that could be generated by biofuels were evaluated in detail, and that remained as central topic to the 2008 FAO report on “The global state of agriculture and food”. The introduction of second-generation biofuels, which use lignocellulosic biomass for the production of bioethanol, opens up a possibility for the production of fuel without competing with food. The lignocellulosic biomass for the production of bioethanol comes mainly from agricultural residue such as pruning remains, stubble, wood remains, corn stover, wheat and rice straw, bagasse (residue from sugarcane and sorghum stalks), etc. The lignocellulosic biomass is the main component of the cell wall of plants, which due to its chemical and structural characteristics is extremely resistant to enzymatic digestion, which is intended to release the sugars trapped in it. So recalcitrant is this structure, that the bioconversion in ethanol of this raw material makes the process currently economically unfeasible. The main objective is that the bioethanol production process becomes economically viable, to achieve this we could do it through a more efficient digestion using more efficient enzymes, by obtaining biomass less recalcitrant and more easily accessible to enzymes, or both. This work has addressed this challenge through a search for lignocellulosic materials more easily accessible to hydrolytic enzymes. For its approach, classical and molecular improvement tools have been used. A collection of sixty-six genotypes of wheat, triticale and barley have been phenotypically characterized throughout their development. The genotypes showed great variability for the saccharification factor, which was negatively correlated with the lignin content in the cell wall. In addition, the results showed that this and other factors could be evaluated with good accuracy using the treatment of images with unmanned aerial platforms (UAV - unmanned aerial vehicle), before harvested, quickly and non-destructively. Moreover, several parents from mapping populations showed contrasting differences for the degree of saccharification. Finally, a tracking of genes involved in the synthesis of wall components was made, and it was observed that the silencing done with miRNA in Brachypodium distachyon produced alterations in the wall cells in the stem, giving rise to, higher degrees of saccharification and alterations the main wall components

Uso de compuestos macrocíclicos derivados de calixarenos como nanotransportadores de fármacos

Los calixarenos son oligómeros cíclicos formados por unidades fenólicas unidas mediante grupos metileno. Estos macrociclos se han convertido en plataformas esenciales para el reconocimiento molecular, la Nanotecnología, la administración de fármacos y la catálisis, debido a su fácil funcionalización tanto en el borde superior como en el inferior de su estructura. La presente Tesis doctoral se centra en mostrar la capacidad de los calixarenos como nanovehículos para transportar material genético y fármacos. Está dividida en seis capítulos. Comienza con una introducción general que cubre varios temas relacionados con el objetivo principal de este trabajo. El Capítulo 2 muestra una descripción amplia sobre los materiales y métodos utilizados para estudiar estos sistemas. Los capítulos 3, 4 y 5 contienen resultados sobre calixarenos de diferentes cargas y propósitos. Finalmente, el Capítulo 6 examina la preparación de liposomas que contienen calixarenos catiónicos anfifílicos y demuestra su potencial uso en terapia génica. En el Capítulo 3, se ha llevado a cabo un estudio de la interacción que muestran de dos calix[4]arenos neutros con ADN de timo de ternera (ctDNA). Los resultados muestran distintos cambios en la conformación del ADN según el valor de X (X= [calixareno]/[ADN]). Los ensayos de citotoxicidad indican que es crucial controlar el diseño estructural de estos sistemas en relación con su biocompatibilidad. El Capítulo 4 estudia la encapsulación del antibiótico antineoplásico doxorrubicina (DOX) en la estructura de un p-sulfonatocalix[6]areno aniónico. Se ha examinado la capacidad de este calixareno para encapsular las moléculas de DOX en su cavidad interna, así como la capacidad de estas moléculas para unirse al ADN. Los ensayos de citotoxicidad muestran una disminución de la toxicidad de la DOX encapsulada en comparación con la DOX libre en diferentes líneas celulares humanas. En el Capítulo 5, se ha investigado la autoagregación de cuatro calix[4]arenos catiónicos anfifílicos en disolución acuosa. Se ha observado la formación de micelas y vesículas, dependiendo del rango de concentración de los calixarenos estudiados. Se han analizado las interacciones entre los agregados de calixareno formados (micelas y vesículas) y el ct-DNA. Los resultados indican que la mayoría de ellos interaccionan con el polinucleótido, invirtiendo su carga y condensándolo, al menos parcialmente. En este capítulo, también se ha investigado la encapsulación de doxorrubicina en los diferentes agregados de calixareno y la posterior liberación de este antibiótico de dichas estructuras. Los resultados demuestran el potencial que presentan las micelas y las vesículas de calixareno como nanotransportadores de fármacos. Finalmente, en el Capítulo 6, se prepararon los liposomas que contienen los calixarenos catiónicos anfifílicos empleados en el capítulo anterior. Se realizó un estudio de las interacciones entre estos liposomas y el ct-DNA. Los resultados muestran que se produce una inversión de carga del polinucleótido, causando una compactación parcial de las moléculas de ADN. Los ensayos de citotoxicidad confirman que estos liposomas presentan una baja y/o moderada toxicidad en las líneas celulares investigadas, por lo que algunos de ellos pueden usarse como nanovectores biocompatibles en terapia génica

The standardisation of vernacular architecture. Wine buildings in Andalusia

[EN] Production buildings constitute a specific section of vernacular architecture, with distinct characteristics. In Andalusia, within this group, the architecture of wine, acquires an important relevancne, the wine cellars. They are a large number of buildings, which were built in the 18th, and 19th centuries. This happened when traditional Andalusian wine production was transformed into a modern wine industry. An industrial development generated a vast architectural ensemble of unique characteristics. This has been studied especially in the Sherry wine region, but it is also present in other regions such as Montilla-Moriles or El Condado de Huelva. The architectural, and industrial wine development in the 19th century was fundamentally based on the repetition of a specific model: the basilica cellar. A simplified formal, and constructive system that comes from the standardisation of the vernacular cellar, and that establishes early points of convergence with the industrial building. A model that continues the tradition in terms of construction, and structure, but conceptually modern in its modular, and repeatable condition. Its reiteration, and extreme simplification made possible the construction of large industrial complexes, and the city transformation. The industrial importance achieved by the wine agro-industry, and the vernacular quality of its architecture introduce different references in Spanish industrial historiography.Research framed within the R&D project "Sistema de Innovación para el Patrimonio de la Andalucía Rural (SIN-PAR)", Code PY20_00298. Andalusian Plan for Research, Development, and Innovation (PAIDI 2020), Junta de Andalucía, European Union, European Regional Development Fund.Aladro-Prieto, J.; Ostos-Prieto, F.; Murillo-Romero, M. (2022). The standardisation of vernacular architecture. Wine buildings in Andalusia. En Proceedings HERITAGE 2022 - International Conference on Vernacular Heritage: Culture, People and Sustainability. Editorial Universitat Politècnica de València. 23-30. https://doi.org/10.4995/HERITAGE2022.2022.15136233

La configuración urbana de Écija en el plano de Manuel Spínola en 1826

Urban cartography in the Spanish 19th century rose during the French occupation and the arrival of the Enlightenment. The maps became instruments that allowed for better city governance. Later, in the reign of Isabel II, the Royal Order of 1846 had as its main objective to establish the control of urban transformations in the main cities of the country through urban cartography. This required a high-plan production by the municipalities themselves, together with the work of cartographers and engineers. In the province of Seville, the cartographer Manuel Spínola de Quintana, worked on seven Andalusian towns, where Écija is one of them. Its plan is possibly the oldest in the city, dating from 1826. It is also the first to represent its urban periphery. It is an unpublished document of great planimetric quality due to its graphics and colouring, as well as its description of the city. A comparison is made with three other contemporary plans of Écija, with a detailed analysis, using Spínola’s plan as the main source. The comparative analysis has produced interesting results, such as a more accurate dating of one of the known plans, as well as evidence of the primacy of Spínola’s document. Finally, a detailed analysis makes it possible to appreciate the configuration of Écija at the beginning of the century, a key factor in future town planning. This is done through the elements of the urban area (parishes, convents, monasteries, squares, etc.), as well as those in its immediate surroundings (roads, farmlands, infrastructures, etc.).La cartografía urbana en el siglo XIX español experimentó un auge durante la ocupación francesa y la llegada de la Ilustración. Los planos pasaron a ser instrumentos que permitían una mejor gobernabilidad de la ciudad. En el reinado de Isabel II, el principal objetivo de la Real Orden de 1846 fue establecer el control de las transformaciones urbanas país a través de la cartografía. Ello demandó un levantamiento auspiciado por los propios municipios, unido al trabajo de cartógrafos e ingenieros. En la provincia de Sevilla, el cartógrafo Manuel Spínola de Quintana dejará constancia de siete poblaciones. Entre estas se encuentra el que es posiblemente el plano más antiguo de la ciudad de Écija, datado en 1826. Además, es el primero en representar su periferia urbana. Se trata de un documento inédito de gran calidad planimétrica debido tanto a su grafismo y cromatismo como su descripción de la ciudad. Se realiza una comparativa con otros tres planos coetáneos de Écija, así como un análisis detallado. Se han generado interesantes resultados como una datación más exacta de uno de los planos conocidos, así como evidenciar la primicia del de Spínola. Un análisis pormenorizado posibilita apreciar la configuración de Écija a principios de siglo, clave en la futura ordenación urbanística. A través de elementos del núcleo urbano (parroquias, conventos, plazas, etc.), así como de aquellos del entorno agreste inmediato (caminos, cultivos, infraestructuras, etc.)

Preparation and Characterization of New Liposomes. Bactericidal Activity of Cefepime Encapsulated into Cationic Liposomes

Cefepime is an antibiotic with a broad spectrum of antimicrobial activity. However, this antibiotic has several side effects and a high degradation rate. For this reason, the preparation and characterization of new liposomes that are able to encapsulate this antibiotic seem to be an important research line in the pharmaceutical industry. Anionic and cationic liposomes were prepared and characterized. All cationic structures contained the same cationic surfactant, N,N,N-triethyl-N-(12-naphthoxydodecyl)ammonium. Results showed a better encapsulation-efficiency percentage (EE%) of cefepime in liposomes with phosphatidylcholine and cholesterol than with 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE). The presence of cholesterol and the quantity of egg-yolk phospholipid in the liposome increased the encapsulation percentage. The bactericidal activity against Escherichia coli of cefepime loaded into liposomes with phosphatidylcholine was measured. The inhibitory zone in an agar plate for free cefepime was similar to that obtained for loaded cefepime. The growth-rate constant of E. coli culture was also measured in working conditions. The liposome without any antibiotic exerted no influence in such a rate constant. All obtained results suggest that PC:CH:12NBr liposomes are biocompatible nanocarriers of cefepime that can be used in bacterial infections against Escherichia coli with high inhibitory activity

Identity construction of the european medium sized city through themonasticism repercussions in Écija

The monasticism appeared during the dawn of Christianity has made a major contribution to Europe's culture and identity. The spread of Christian faith throughout the continent was achieved not only through a new morality and religion but also through art and convent architecture. The Middle Ages were the peak of this monastic production. Orders such as Cistercians or Dominicans would appear mainly in France and Italy spreading their rules and ways of life to the whole continent. It is a phenomenon that transcends borders, kingdoms and geographical elements, bringing together many people under the same identity.The flag of the Latin cross waved in the European Middle Ages. However, the reality in southern Iberian Peninsula was different. Al-Andalus stood as a bastion of Islam in Europe long before the appearance of the Ottoman Empire. At the end of the Middle Ages, the Kingdom of Castile began to occupy territories held by the Muslim Kingdom of Granada. The need to Christianisethe new conquered lands would call a multitude of monastic orders settled in Europe to focus their attention on the new Kingdom of Seville. The main cities such as Seville, Cordoba, etc. would be the most coveted for the new foundations. Nevertheless, Écija is located in the Guadalquivir river valley in a difficult position on the border with the Kingdom of Granada. In the 14th Century, Écija had close to 10,000 inhabitants, which, at the time, was a large number. A city which was an attraction point for Christian Communities, which according to their origin, 31% came from Italy, 22% from France, 37% from Spain and 10% from Jerusalem. The study of Écija shows the crossing of continental relationships through monasticism. How could the influence of Christianity manage to cover an entire continent to the most remote place? We could find the keys to understand the urban construction of western monasticism through the main average conventual cities in Andalusia

Binding of DNA by a dinitro-diester calix[4]arene: Denaturation and condensation of DNA

A study of a dinitro-diester calix[4]arene (5,17-(3-nitrobenzylideneamino)-11,23-di-tert-butyl-25,27-diethoxycarbonyl methyleneoxy-26,28-dihydroxycalix[4]arene) interaction with calf-thymus DNA was carried out using several techniques. The measurements were done at various molar ratios X=[calixarene]/[DNA]. Results show diverse changes in the DNA conformation depending on the X value. Thus, at low macrocycle concentrations, the calixarene binds to the polynucleotide. This interaction, mainly in groove mode, weakens the hydrogen bonds between base pairs of the helix inducing denaturation of the double strands, as well as condensation of the macromolecule, from an extended coil state to a globular state. An opposite effect is observed at X molar ratios higher than 0.07. The de-condensation of DNA happens, that is, the transition from a compact state to a more extended conformation, probably due to the stacking of calixarene molecules in the solution. Results also show the importance of making a proper choice of the system under consideration.Junta de Andalucía P08-FQM-03623, P12-FQM-110

Assessment of the denaturation of collagen protein concentrates using different techniques

The use of collagen and gelatin in the field of regenerative medicine is widely extended. However, most of the studies in this topic are focused on the scaffolds’ properties, but only a few are related to the properties of the raw material used. The raw material analysis not only consists of a study of the composition, but also of the denaturation degree that can influence the processing and properties of the structure of the scaffold. Thus, the denaturation degree analysis of different collagen proteins was performed and assessed by the comparison of four different methods: differential scanning calorimetry (DSC), Fourier transform Infrared Spectroscopy (FTIR) and circular dichroism (CD) spectra and sulfhydryls content analysis. DSC measurements put forward a glass transition between 88°C and 95°C as well as from the FTIR measurements; the characteristic peaks for proteins are evidenced. However, from the sulfur content, only a small proportion of free sulfhydryls are present with respect to their total amount. In addition, CD spectra allow to estimate the secondary structure of the protein by the analysis of the α-helix and β-strand and also quantify the denaturation degree with the ‘positive/negative ratio’ (RPN) from the CD profiles, obtaining values in the range between 25% and 100%.Ministerio de Economía y Competitividad (Spanish Government) / MINECO/FEDER (UE) CTQ2015-71164-

Potentiometric study of carbon nanotube/surfactant interactions by ion-selective electrodes. Driving forces in the adsorption and dispersion processes

The interaction (adsorption process) of commercial ionic surfactants with non-functionalized and functionalized carbon nanotubes (CNTs) has been studied by potentiometric measurements based on the use of ion-selective electrodes. The goal of this work was to investigate the role of the CNTs' charge and structure in the CNT/surfactant interactions. Non-functionalized single- (SWCNT) and multi-walled carbon nanotubes (MWCNT), and amine functionalized SWCNT were used. The influence of the surfactant architecture on the CNT/surfactant interactions was also studied. Surfactants with different charge and hydrophobic tail length (sodium dodecyl sulfate (SDS), octyltrimethyl ammonium bromide (OTAB), dodecyltrimethyl ammonium bromide (DoTAB) and hexadecyltrimethyl ammonium bromide (CTAB)) were studied. According to the results, the adsorption process shows a cooperative character, with the hydrophobic interaction contribution playing a key role. This is made evident by the correlation between the free surfactant concentration (at a fixed [CNT]) and the critical micellar concentration, cmc, found for all the CNTs and surfactants investigated. The electrostatic interactions mainly determine the CNT dispersion, although hydrophobic interactions also contribute to this process.Fil: Ostos, Francisco José. Universidad de Sevilla; EspañaFil: Lebrón, José Antonio. Universidad de Sevilla; EspañaFil: Moyá, María Luisa. Universidad de Sevilla; EspañaFil: Bernal, Eva. Universidad de Sevilla; EspañaFil: Flores, Ana. Universidad de Huelva; EspañaFil: Lépori, Cristian Marcelo Oscar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Física Enrique Gaviola. Universidad Nacional de Córdoba. Instituto de Física Enrique Gaviola; ArgentinaFil: Maestre, Ángeles. Universidad de Huelva; EspañaFil: Sánchez, Francisco. Universidad de Sevilla; EspañaFil: López Cornejo, Pilar. Universidad de Sevilla; EspañaFil: López López, Manuel Carlos. Universidad de Huelva; Españ

Heritage replacements: from convent to square and to contemporary architecture, Conventual urban transformations in Andalusian cities

In Andalusia (Spain), the conventual typology has shaped the urban centers of its cities following a process of implementation, which affects the traffic and internal organization of the city. In the mid-19th century, the confiscations led to the expropriation, demolition, and disappearance of a large part of their monastic spaces. The starting situations (size of cities, socioeconomic state in the 19th century or characteristics of the conventual foundations) gave rise to diverse urban responses. The bourgeoisie's need to "modernize" the cities led to the appearance of squares and the opening of new roads. On other occasions, the old factories were reused for the installation of new uses (prisons, barracks, markets, etc.), or their plots were used for new construction. Τhe case of Plaza Nueva in Seville from the demolition of the San Francisco Convent is studied. Also, together with other Andalusian examples, such as the San Antonio de Padua Convent in El Puerto, the current Plaza Isaac Peral and Los Descalzos Convent in Écija. The case of this last city constitutes the counterpoint in the convent reuse, without generating relevant urban spaces. Once again, the Plaza Nueva, due to its condition of the centrality of the Andalusian capital, constitutes the maximum exponent of this urban and symbolic revision. The economic power put into practice its urban capacity, again through architecture. It will finance new buildings as representative images of their brands, companies, and institutions. The testimonies of the convent activity, the new spaces emerged from the disentailment actions and their new contemporary symbols constitute sequential fragments of urban history. They are necessary for the cities for their valuation and heritage understanding. The study of these Andalusian cases can serve as a reference for the detection of similar processes in the European Mediterranean frame